BRB 747A, 2109 Adelbert Road, Cleveland, Ohio 44106
Dye-terminator sequencing is performed using a modified protocol supplied by Applied Biosystems BigDye Terminator 1.1 and 3.1 Cycle sequencing kits. All reactions are analyzed using a 48-cappilary 3730 Genetic Analyzer. Sanger Sequencing can be performed on a variety of templates including PCR product, plasmids, and BACs for analysis of mutation detection and traditional DNA sequencing.
The Genomics Core offers quality sequencing data, with a fast turn-around time, and knowledgeable help with troubleshooting. Discounts are available for large projects.
|Type of Template||Quantity Required|
|Plasmid||100-300ng per reaction|
|Short PCR product (<500bp)||50-100ng per reaction|
|Long PCR product (>500bp)||100-200ng per reaction|
|Primer||2ul of 20uM concentration per reaction|
Universal primers stocked by the Core
|BGH reverse||5’ TAGAAGGCACAGTCGAGG|
|M13 Forward||5’ GTTTTCCCAGTCACGAC|
|M13 Reverse||5’ AACAGCTATGACCATG|
|T7 promoter||5’ TAATACGACTCACTATAGGG|
Data can be accessed through the Helix fileserver which is accessible to all systems on the Case network. An account for your lab can be created where data files can be securely stored and retrieved. Please email the Genetics Department Computer Systems Administrator, Ulrich Neuss (Uli) at firstname.lastname@example.org to set up an account. In the e-mail please specify your name, e-mail address, your principal investigator, and explain that you would like access to the facility to get your data.